DE1793269C3 - Process for the production of saponins and sapogenins - Google Patents

Description

The invention relates to the extraction of steroid Sapophins and sapogenins from vegetable Material-

Plant material that provides sapogenins grows in fallen parts of the world. The ones obtained from them Sapogenins have considerable economic benefits; · Ο Importance. They can be used as starting materials for the production of various biologically active sieroidiicher Use compounds, especially for the manufacture of corticosteroids ^ To the most economical ι ο Important sapogenins include diosgenin, kecogenin, and the sapogenins from Solanum, all of which are steroid Are alkaloids. Sapogenins are usually found in plant material in the form of saponins relatively small amounts of, for example, about 2%. Much work has been done on Processors to extract them effectively. There are already various types of pretreatment of the plant material has been provided to increase or decrease the amount of sapogenin available make the process more effective. These processes include, for example, fermentation of the plant material prior to extraction or mechanical grinding of the material to make the cell walls to break as much as possible.

It has now been found that the total amount of sapogenin to be obtained can be increased if the vegetable material from which the sapogenin is to be extracted before extraction of the sapogenin is treated with certain substances that influence the plant metabolism. It is not exactly known which conversions take place here. Perhaps these substances promote biosynthesis of sapogenin supplying material, e.g. B. by promoting enzyme activity, or perhaps preventing it Effectiveness of enzymes that break down the sapogenin-producing material.

The invention relates to a method for obtaining steroidal saponins or sapogenins from The process is characterized in that one the plant material before the extraction of the saponins or sapogenins with a regulator that changes normal plant metabolism or growth. at Using the method, increased yields of steroidal saponins or sapogenins are obtained in comparison with otherwise similar procedures, but without the use of a regulator.

It has not yet been possible to link the effectiveness of the regulator to certain chemical structures. After all, the effective regulators to be used according to the invention, which are usually organic compounds, belong to a relatively small number of groups with recognized biological activities. These groups are:

(a) natural regulators that affect plant growth, including auxins and hormones,

(b) synthetic regulators that affect plant growth, including auxins, hormones and herbicides,

(c) sulphydryl inhibitors,

(d) regulators of lipid metabolism, <> 5

(e) regulators of steroid metabolism,

(f) naturally occurring and racemic α-amino acids,

(g) Group B vitamins, rutin and water soluble

Derivatives of vitamin A and tocopherol,
(h) Analogs of growth factors, such as vitamin additions

timetableabolite and amino acid antimetabolites,
(1) Antibiotics from the class of penicillins, the

Griseofulvine and the chloramphenicol.
The regulators to be used according to the invention can also have more than one of the abovementioned properties and can therefore also be classified in more than one of classes (a) to (i). Mixtures of such substances can also be used. These mixtures are discussed in more detail below.

There are, of course, significant differences in the biological. Activities of these classes. On off The system consisting of several enzymes works within the plant material, which is the sapogenin supplies. The regulator present can act directly by promoting the biosynthesis of saponin or by preventing the breakdown of saponin. But the effect can also be more indirect Ways to be reached. For example, the regulator does not need to be a regulator for the enzyme itself but can be chemically or through the action of endogenous enzymes or another regulator physically transferred to a regulator. In some cases, the regulator can also act in such a way that that it does not promote the formation or the release of the sapogenin, but on certain processes acts within the cells, which in the end result in an increase in the yield of sapogenin is achieved.

Some of the regulators to be used according to the invention are mentioned below. This list, for example, is not intended to limit the scope of the invention.
Class (a)

Naturally occurring regulators that influence plant growth: 3-indole acetic acid, gibberellins, kinetin (6-furfurylaminopurine), trauma acid, abscissic acid.
Class B)

Synthetic regulators that influence plant growth: derivatives of acetic acid, propionic acid, Butyric acid, maleic acid, acrylic acid, benzoic acid and picolinic acid, for example 2,4-dichlorophenoxyacetic acid and other chlorinated Phenoxyalkansäureri, also their salts, esters unc Amides,

Trichloroacetic acid, 4-amino-3,5,6-trichloropicolinic acid;

Derivatives of naphthalene, e.g. B. methyl 1-naphthalene acetate;

Bipyridylium herbicides, e.g. B. l.l'-ethylene-2,2'-bipyridylium dibromide and
1,1 '-DimethyM ^' - bipyridylium dichloride;
Carbamates, ζ. B. Isopropyl N- (3-chlorophenyl) carbamate;

Amides, ζ. B. 2-chloro-N, N-diailyl-acetamide;
Urea compounds, ζ. B. N '- (4-chlorophenyl) -N, N-dimethylurea;
Diazines, ζ. B. 1,23,6-tetrahydro-3,6-dioxopyridazine;

Triazines, ζ. B. 2-chloro-4-ethylamino-6-isopropylamino-133-triazine;
Phenols, e.g. B. 2- {1-methylpropyl) -4,6-dinitrophenol;

substituted uracils, ζ. B. 5-bromo-3-sec-butyl-6-melhyluracil;

various herbicides such as ethylene glycol bis-trichloroacetate;

2,3,6-trichlorobenzylo, propanol; 2,6-dichlorobenzonitrile; 3-amino-1,2,4-triazole;
2,6-dichloro-thiobenzamide;
Disulphides such as di (ethoxythiocarbonyl) disulphide;

organic arsenic compounds, ζ. Β. ι ο

Disodium methane arsonate;
N-6-benzyladenine,
Class (c)

Sulphydryl inhibitors: derivatives of lactic acid and maleic acid, indoles, for example N-ethylmalemide, N-phenylmalemide, DL-3-indolactic acid, 5-hydroxyindole, 3-methylindole; Iodoacetic acid, iodosobenzoic acid and iodoacetane / id.
Class (d)

Regulators of lipid metabolism: The enzyme preparation Vasolastine (Enzypharm CV).
Class (s)

Steroid Metabolism Regulators:
Ethyl a- (4-chlorophenoxy) -a-methyl propionate;
2- (p-chlorophenyl) -1 - [pf 2- (diethyl-

amino) ethoxy] -phenyl] -lp-tolyl-ethanoI;
Lipostabil, a preparation made from soybeans.
Class (noun)

Naturally occurring and racemic amino acids: L-Valine, L-Threonine, DL-Isoleucine, L-Leuein.
Class (g)

Group B vitamins: choline chloride, pyridoxal hydrochloride, riboflavin, thiamine hydrochloride, vitamin growth factors such as preparations made from yeast, cottonseed, cereals and soybeans. Such preparations usually also contain amino acids.
Class (h)

«-Picolinic acid hydrochloride, DL-desthiobiotin, AI-lyl-DL-glycine.
Class (i)

Antibiotics: The sodium salt of penicillin G, chloramphenicol, griseofulvin.

Following the process step according to the invention, the sapogenins can be obtained directly from the plant material by known methods, e.g. B. by treating the treated vegetable material with a substance which, like an aqueous acid or water, can hydrolyze the glycosidic bonds under pressure. Then you win the sapogenins from the hydrolysis product, z. B. by extraction with a solvent. However, the sapogenins can also be obtained indirectly from the plant material by extracting the treated plant material with a solvent for saponin such as methanol. The glycosidic bonds in the extract are then hydrolyzed with an acid to release the sapogenins. These can then be obtained from the hydrolyzate, for example by extraction with a solvent. In both ways, treatment with the regulator should be carried out prior to hydrolysis.

The method according to the invention relates in particular to the extraction of steroid substances from the material of monocotyledons, which are already used as raw material for the extraction of sapogenins , e.g. B. the fruits, leaves, sprouts, rhizomes, tubers or roots of Dioscoreacees, Liliai-een and Amaryllidaceae. These include in particular the various Dioscorea sylvatica, Dioscorea composita, Dioscorea floribunda, Dioscorea mexicana, Dioscorea tokoro, Dioscorea deltoidea and other species such as Tamus, Agave, Aleiris, Asparagus, Similax, Trillium. You can also use the process according to the invention to obtain sapogenins from dicotyledons, e.g. B. from legumes, Solanaceae, Balanitaceae and Zygophyllaceen, in particular from the seeds and fruits of Trigonella, Trifolium, Solanum and Balanites, such as Trigonella foenumgraecum, Trigonella coerulea, Trigonella corniculata, Trigonella cretianum, Trifolium ornithopodiodesatum, Solananum hasasatum. Balanites aegyptiaca (Balanites roxburghii), Balanites orbicularis, Balanites pedicellaris and Balanites wilsoniana.

It has been proposed to incubate plant sapogenin-producing material by holding it in an aqueous medium for a suitable period of time, often at an elevated temperature. If necessary, water is added to the plant material in such an amount that there is a separate aqueous phase during the entire incubation period. This pretreatment can advantageously be included in the method according to the invention. When doing this incubation, e.g. B. during a period of 1 to 30 days, and in the presence of a regulator, which is added at the beginning or during the incubation period, the yield of sapogenins can be increased. Part of the increased yield can probably be explained by the incubation itself. However, comparative tests have shown that the increased yield is due to a substantial extent to the presence of the regulator. Even if the regulator is added shortly before the steroid material is obtained, ie shortly before hydrolysis, the amount of sapogenin that can be obtained is increased compared to control extractions without a regulator. In this embodiment of the method, the vegetable material can be incubated in the presence of an aqueous medium, for example for 1 to 30 days, then treated with the regulator and hydrolyzed. Of course, the incubation is not absolutely necessary.

As already noted, the regulator influences the biosynthesis of the sapogenin-producing material and causes an increased yield. However , the regulator probably does not take part in the biosynthesis itself. It is therefore only necessary to add small amounts of the regulator, usually not more than about 1000 parts by weight per million parts by weight of air-dry plant material with a water content of not more than 10%. Typical amounts are 50 to 100 parts by weight of regulator per million parts by weight of air-dry plant material.

The regulator can be used as a pure chemical substance or be added in the form of mixtures containing the regulator as an active substance.

Plant material to be treated in the form of tubers can be crushed before treatment, so that the liquids have free access to all parts. Tubers can be z. B. too thin Cut up slices, and then chop up so that the cell walls are broken. You can see the slices

also dry and then powder. A pretreatment can also consist in the fact that the vegetable Material incubated with degrading enzymes such as cellulase or pectinase. This will make the regulator and later brought the hydrolyzing agent into contact with as large a number of cells as possible. If a pretreatment with an enzyme is used, this pretreatment and the one mentioned above are used Incubation is best done at the same time. The regulator can be used at any time during added to the incubation period.

The incubation in connection with the method according to the invention is preferably carried out in such a way that a sufficient amount of water is added to the plant material so that a special aqueous phase is present at the end of the incubation period. Additional water is usually required when treating seeds and dried tubers and leaves. The mixture is preferably maintained at an elevated temperature, for example from 25 to 4O ⁰ C or up to 8O ⁰ C, in the presence of added regulator. The yield of sapogenins depends on the incubation time and the incubation temperature. During the incubation, the steroid material is continuously biosynthesized and continuously broken down. Simple experiments can be carried out to determine the optimal incubation times and temperatures

will. ,,.

The extraction of the sapogenins from the treated product can be done by doing it with a Mineral acid treated, whereby the glycos.dischen bonds are hydrolyzed. One can for example reflux an incubation product with 2N hydrochloric acid for 5 hours during which the acid-insoluble sapogenins are released. Instead of hydrochloric acid, other non-oxidizing agents can also be used Mineral acids, such as sulfuric acid, can be used, as well as other acids or acidic salts. The material insoluble in the acids is then separated from the hydrolyzate, e.g. B. by filtering or Hurl off. The sapogenins can then be extracted from the acid-free, insoluble cuckoo with e.nem Solvents such as petroleum ether can be extracted.

Using a material from dicotyledons, e.g. B. seeds, they should not be crushed too much, as described for example in British patent Π 36,626. However, it may be appropriate to crush larger seeds, for example of balanites, into smaller pieces, seeds containing oils, e.g. B. from Balanites, are preferably at least partially degreased before adding the regulator. . ,. a

For the production of the starting material, the vegetable material can be used according to the method of the British Patent 1198 626 with certain saturated Hydrocarbons, which are likely to take part in the metabolism, are pretreated.

The saturated hydrocarbons to be used here are those with 10 to 36 carbon atoms in the molecule. These include, for example, n-hexadecane, n-Eicosan, n-Pentacosan, n-Hexacosan and n-Uctacosan. Likewise, branched chain hydrocarbons can also be used, e.g. B squalane (2,6,10,15,19,23-hexamethyltetracosane). Similar success <vs. ge can be achieved using the regulator with precursors of steroids. e.g. mevalonic acid or its derivatives or squalene.

Perhaps the particularly high yields of sapogenin can be explained by the fact that the imported saturated hydrocarbons or the precursors of a steroid during incubation sapogenins biosynthesize. Perhaps these substances take part directly or indirectly in biosynthesis. The presence the regulator reduces the loss of sapogenin.

The method according to the invention can be used with particular success in the extraction of sapogenins such as diosgenin from plants of the genera Dioscorea, Trigonella and Balanites. The examples below explain the procedure. The quantities given by the regulator relate to the weight of the vegetable material in the state in which it was used. This is an on air dried material that contains less than 10% water. The yields are given in relation to anhydrous, vegetable material.

example 1

Treating Dioscorea deltoidea with indole-3-acetic acid (IAA) as regulator.

G.I. Process A Acid hydrolysis alone

100 g of dried and crushed tubers of Dioscorea deltoidea with particle sizes below 2 mm and a moisture content of 5.5% were refluxed with 730 ml of 2N hydrochloric acid in a conical bottle with a capacity of 2 1/2 hours. The mixture was then cooled to room temperature, the acid-insoluble material was collected, washed with water and then with a 5% ammonia solution until it became alkaline. The insoluble residue was dried for 16 hours at 6O ⁰ C and crushed him in a hand grinder to such an extent that all the particles had a-diameter of less than 1.7 mm. This powder was extracted in a Soxhlet apparatus for 48 hours with 1200 ml of light petroleum having a boiling range of from 40 to 6O ⁰ C. The petroleum extract was allowed to stand at room temperature for 16 hours. The crystalline diosgenin was filtered off and washed with light petroleum, a first portion of white crystals being obtained. The mother liquor and the wash water were concentrated to a volume of 200 ml. This produced a second portion of white crystals.

G. 2. Method B / b Acid hydrolysis with a regulator

2.5 ml of a solution of indole-3-acetic acid (48 mg in 10 ml of absolute alcohol) was poured into a 2 liter conical bottle pipetted in. It was allowed to evaporate overnight. 100 g powdered was put into the bottle Tubers and 730 ml of 2N hydrochloric acid. The treatment then became just as long and just as long continued as described above under G. 1..

G. 3. Method C Incubation without regulator

100 g of the vegetable material described above was placed in a 2 liter bottle. 600 ml of tap water were added and the mixture was shaken

709 646/49

Minutes. The bottle was then placed in a dark incubator and left at 37 ° C for 48 hours itehen. The bottle was shaken 2 hours after the introduction, and also after more Hours. Then 130 ml of concentrated hydrochloric acid was added closed, refluxed and worked up in the same way as described under G. 1..

G. 4. Method C Incubation with regulator

As described under G. 2., the regulator was placed in a 2 liter bottle. The next Tomorrow the powdered tubers were added so that the incubation began at the same time as with G. 3. The both experiments were then carried out together.

10 The concentrations of the regulator and the Incubationszeiten are given in Table I. The yields indicate the amounts by weight of sapogenin that was actually obtained as a solid material.

Table I.

Example procedure

Regulator incubation time Yield in g

1st ant. 2nd ant.

Melting point, ⁰ C 1st ant. 2nd ant.

Total sapogenin yield

p.p.m.

hours

B / b

O 20

0 20

48

48

3.11 0.82 210 206-7 4.16 3.43 0.63 209 207-8 4.30 4.21 0.54 210 205-6 5.02 4.00 1.04 210 206-7 5.33

A comparison of G 2 with G 1 and of G 4 with G shows that the regulator has a beneficial effect.

The use of IAA as a regulator was tested in further experiments, with the amounts of IAA and the duration of the incubations were as follows:

Procedure C was modified to procedure D / a, the incubation being carried out without a regulator, but the regulator being added after the hydrochloric acid and before the hydrolysis. In experiment D / b, the incubation was carried out without a regulator, and the regulator was added before the addition of the hydrogen chloride and before the acidic hydrolysis. In experiments 1-16, 2.5 g each of dried tubers of Dioscorea deltoidea 40 were used

placed in a bottle that may contain IAA. The IAA was made as a solution in ethanol added, whereupon the solvent was removed. Then 25 ml of tap water were added each time, shook the mixture for 5 minutes, washed the walls of the bottle with another 25 ml of tap water and placed the bottle in a dark incubator maintained at 37 ° C. Then the product was hydrolyzed and extracting the acid insoluble material as described in Method A. One obtainable raw diosgenin. This product was then examined by thin layer chromatography, whereby mar used a Chromoscan densitometer. The results are shown in Table II.

Table II

Attempt no.

proceedings

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

B / b

D / b

There

Regulator Incubation period Chromoscan concentration Sapogenin p.p.m. hours Yield in% 0 0 4.20 200 0 4.31 0 24 4.68 0 48 4.65 0 72 4.68 2 24 4.98 2 48 5.18 2 72 5.24 20th 24 5.18 20th 48 5.41 20th 72 5.21 200 24 5.31 200 48 5.40 200 72 5.24 200 48 5.55 200 48 5.43

A comparison of the results shows you the beneficial effect of the regulator.

example

Treatment of Dioscorea deltoidea with Giberellin (A + X)

The procedures described in Example 1 were repeated on the same scale, using as regulator Giberellin (A + X) used, made by Mann

Table III

Research Laboratories Inc. was manufactured. The results are given in Tables III and IV.

attempt

Procedure regulator incubation period

Yield in g

1st part 2nd part Melting point, "C 1st part 2nd part

p.p.m.

Hours total yield of sapogenin Vo

G5 A. 0 0 2.83 0.86 209 206-7 3.91 G6 B / b 20th 0 3.68 0.07 210 197-8 3.97 G7 A. 0 72 3.58 0.99 210 207-8 4.84 G8 C. 20th 72 3.32 1.66 210 207-8 5.29

A comparison of G 6 with G 5 and of G 8 with G 7 shows the beneficial effect of the regulator.

Table IV

attempt

No.

proceedings

Regulator concentration p.p.m.

incubation

length of time

hours

Yield of sapogenin

1 A. 0 0 4.20 2 B / b 200 0 4.52 3 C. 0 24 4.68 4th C. 0 48 4.65 5 C. 0 72 4.68 6th C. 2 24 5.00 7th C. 2 48 5.30 8th C. 2 72 5.18 9 C. 20th 24 4.91 10 C. 20th 48 5.42 11th C. 20th 72 5.86 12th C. 200 24 5.18 13th C. 200 48 4.98 14th C. 200 72 5.17 15th D / b 200 48 5.05 16 There 200 48 5.05

These tests also show the beneficial effect of the regulator.

Example 3 Treatment of Trigonella foenumgraecum with indole-3-acetic acid (IAA)

5 g of whole seeds of Trigonella foenumgraecum were added to 25 ml of tap water. One incubierte the mixture in the dark for 6 or 24 hours at 37 ⁰ C. Then was added to hydrochloric acid, so that a 2normale solution. The mixture was refluxed for 2 hours. After cooling, the mixture was filtered off, the acid-insoluble residue was washed with water and dilute ammonia until all traces of acid had been removed, dried and extracted with Pctrolether in a Soxhlet apparatus. The organic solvent was then evaporated leaving a residue of crude diosgenin. Various amounts of IAA were added to the mixture before placing it in the incubator. A comparison was carried out without incubation, with the seeds immediately being acidic hydrolyzed. The yields were determined by examining the crude product in chloroform using an infrared spectrophotometer.

Table V

Sapogenin yield in%

Ineubation stealer, style. 0 6 24

Trigonella alone 1.13

Trigonella + 1 ppim. IAA
Trigonclla -f 10 ppm IAA
Trigonella + 100 ppm IAA
Trigonella +800 ppm IAA 1.11

1.06 , 01 1.33 , 37 1.26 , 34 1.29 , 34 , 14 ,If

Example 4
Treatment of Trigonella foenumgraecum

with Gibbereilinen

The procedures described in Example 3 were repeated using a 10% solution of the Potassium salt of gibberellic acid used in place of IAA. The results are in Tables VI and VH included.

Table VI

Gibberellin (A + X) p.p.m. Yield of sapogenin ion duration. Hours. in % 6th 24 p.p.m. Incubat! 1.06 1.01 0 1.28 133 Trigonella alone p.p.m. 1.13 Trigonella + 1 1.09 13th Regulator Trigonella + 10 1.14 1.29 Regulator Trigonella +100 Regulator Table VII

Acid potassium salt of the gibberellin yield of sapogenic acid (10%) in%

Incubation time, hours
0 6 24

Trigonella alone U3 1.06 1.01 Trigonella + 1 p.p.m. 1.28 13th Regulator Trigonella + 10 p.p.m. 1.10 1.19 Regulator Trigonella + 100 p.p.m. 1.19 134 Regulator

Example 5

Procedure C according to Example 1 was repeated for the treatment of Dioscorea deltoidea, Trigonella foenumgraecum or Balanites aegyptica with different regulators and different incubation periods at 37 ° C. The results are given in columns D, F and B of the tables below. After the incubation, the material became acidic

ίο hydrolyzed, as in method A of example 1 Unless otherwise stated, the content of the sapogenins in the hydrolyzate is described detected by infrared spectroscopy. * means the results obtained with the Chromoscan device according to Example 1 are obtained. The yield is given in percent, based on the air-dry Source material. The figures given in the "Percentage increase" column mean the increase in Yield compared to a yield under the same conditions but with no regulator.

The experiments with Dioscorea were carried out with tubers that were air dried and then were crushed. Except where otherwise stated the experiments with Trigonella were carried out with whole seeds. The sign "+" means that the crushed seeds were partially defatted. Unless otherwise stated, the experiments were performed with balanites with crushed seeds that were partially defatted (the remaining oil content was 3-20% by weight).

Examples 1 to 4 describe the use of naturally occurring plant growth regulators of group (a).
The use of substances from groups (b) to (i)

is described in the tables below.

Regulator

concentration
pp. in.

Incubation period
hours

Increase in
DF

Pharmamedia 105 6th 5 28 Proflo 105 48 Yeatex granules ΙΟ * 6th 8th 39 Yeatex great 10s 48 Lipostable 5x10 « 24 48 Vasolastins 2x105 24 42

Pharmamedia is a preparation from cottonseed germs, the main ingredient being a non-hydrolyzed globular protein

Yeaiex is a spray-dried, aqueous yeast extract Lipostable is a mixture of certain fractions of soybean phosphatides and pyridoxine hydrochloride in a weight ratio of 400: 3. Vasolastin is obtained from ampoules that contain a Mixture of 8,000 units of lipid metabolism enzymes, 4,000 units of tyrosine, 4,000 Amine oxidation units and a stabilizing solvent contain

Proflo is partially defatted boiled cottonseed meal.

Regulator

Ethyl - * - (4-chk> rphenoxy) -

Λ-methyl propionate

Ethyl - «- {chlorophynoxy) ·

Λ-methyl propionate

Ethyl - «- (4-chlorophenoxy} -

Λ-methyl propionate

concentration Incubation period Increase in 34 p.pm hours D 1- 10 ' 48 6th 5x10 « 24 5x10 « 24

continuation 17 93 269 Incubation period 16 6th F. H B. 8th 7th 15th Regulator hours 6th 24 6th 10 8th Alloxane monohydrate concentration 24 Alloxane monohydrate p.p.m. 48 5 15th N-6-benzyl-adenine 400 24 Increase in% 5 2,4-dichlorophenoxy acetic acid 400 24 D. 2,4,5-trichlorophenoxy acetic acid 400 12th 8th 2,4,5-trichlorophenoxy-propion- 400 8th acid 400 CTl i4 3-indole butyric acid 10.8 72 Iodoacetic acid 24 9 CTl 13f Iodoacetic acid 400 48 7th Iodoacetamide 400 6th 9 + 6th Iodosobenzoic acid 400 72 «-Naphthalene-acetic acid 400 6th 16 «-Naphthalene-acetic acid 400 24 α-naphthalene acetic acid 400 72 10 7 + N-ethyl maleimide 400 24 Maleic hydrazide 400 CTi 5 8th-/- Maleic hydrazide 400 24 / J- (2-furyl) acrylic acid 400 CTl 12th ] 3- (2-furyl) acrylic acid 400 24 5 5 l, l'-dimethyl-4,4'-dipyridylium- 400 24 CTl 13th Nicotinic acid amide 400 24 11th Pyrodoxine hydrochloride 1.6x103 CTl 6th 6th Riboflavin 400 24 8th Thiamine hydrochloride 400 24 15th D-calcium pantothenate 400 24 7th Choline chloride 400 24 15th Acetylcholine chloride 400 24 7th Folic acid 400 24 15th Meso-inositol 400 24 11th Nicotinic acid 400 24 14th Pyridoxal hydrochloride 400 24 12th p-aminobenzoic acid 400 6th 8th DL-α-tocopherol acetate 400 CTl 13th Rutin 400 6th CTl Vitamin A, acetate 6x103 24 14th DL ethionine 400 24 19th DL-3-thienyl-DL-alanine 400 6th 9 Glycine 400 6th 14th Allyl-DL-glycine 400 CTl 8th DL-0-phenyllactic acid 400 24 15th «-Picolinic acid hydrochloride tOO 24 14th 2-chloro-4-aminobenzoic acid 400 24 11th Oxythiamine hydrochloride 400 24 13th DL-desthiobiotin 400 CTl 17th Deoxypyridoxine hydrochloride 400 24 Chlcramphenicol 400 24 Propranol hydrochloride 400 24 K.erbisan-5 [diethyldithio- 2XlO ⁴ bis (thionoformat), 58%] 2x103 24 5-bromo-6-methyl-3- (l -methyl- 4x103 propyl) uracil 48 Griseofulvin 400 48 Penicillin G, sodium salt 5x10 « 5 χ 1 (H A units per g 48 Orotic acid (uracil-4-carboxy- air drier acid Tubers 48 3-amino-1,2,4-triasol 400 48 DL isoleucine 400 4x103

example

18th

The procedure and calculations of Example 5 were repeated with the difference that instead of a single regulator mixtures of two regulators or one or more regulators along with a precursor of a steroid or a long chain saturated hydrocarbon called the acting as a precursor to a steroid was used. The yield increases are based on comparable ones Try without additives.

additions

(a) DL-mevalonic acid lactone
2,4,5-trichlorophenoxy-propionic acid DL-mevalonic acid lactone
together with 2,4,5-trichlorophenoxypropionic acid,

both added at the beginning of the incubation

(b) 2,6,10,15,19,23-hexamethyl-tetracosa-1,5x105 2,6,10,14,18,22-hexaen (squalene) gibberellic acid (90% +) 138 Squalene 1.5x105

along with gibberellic acid (90% +), 138 both added at the beginning of the Incubation

(c) n-Hexadecane 2xl0 ^ä \ together with gibberellin (A + X), 480 j, both added at the beginning of the incubation

(d) n-Hexadecane 2xl0 ⁵ j together with gibberellin (A + X) and 480 \ 3-indole acetic acid, 480 J all added at the beginning of the incubation n-hexadecane, 2xlO ⁵ added at the beginning of the incubation 430 together with gibberellin (A + X)

and 3-indole acetic acid, 480

both added 24 hours after incubation

concentration Incubation period p.p.m. hours 2.5 x 10 ⁴ 6th 1.08 6th 2.5x10 ^ I 6th 1.08 y

Yield increase in% DFB

12 12

24

24

72

36

26th

Claims (22)

Patent claims: 1. Process for the production of steroidal saponins or sapogep.inen from stereoidal ones Plant material supplying sapogenins, thereby characterized in that the vegetable material before the extraction of the saponins or treat sapogenins with a substance that normal plant metabolism or the norma- ι ο Ie influences plant growth (so-called regulator) and one of the following groups of substances belongs to: (a) natural regulators that influence plant growth, (b) synthetic regulators that influence plant growth, (c) sulphydryl inhibitors, (d) regulators of lipid metabolism, (e) regulators of the steroid metabolism,> o (f) naturally occurring and racemic "amino acids, (g) vitamins of group B, rutin and water-soluble derivatives of vitamin A and tocopherol, (h) analogs of growth factors such as vitamin antimetabolites and amino acid antimetabolites,
(i) antibiotics from the class of penicillins, the Griseofulvine and the chloramphenicol
and the process is carried out with regulators which acidic hydrolyze the saponin in such a way that the yield increases without the saponin being hydrolyzed. 2. The method according to claim 1, characterized in that the regulator is indole-3-acetic acid or a Giberellin used. 3. The method according to claim I _1, characterized in that a chlorinated phenoxyalkanoic acid or a salt, an ester or an amide of such an acid is used as a regulator; Indole-3-butyric acid; Naphthalene-acetic acid or a salt, an ester or an amide of this acid; l, r-dimethyl-4,4'-dipyridylium dibromide; j3- (2-furyl) acrylic acid or 3-amino-1,2,4-triazole are used. 4. The method according to claim 1, characterized in that that one uses a mixture of enzymes that regulate the lipid metabolism. 5. The method according to claim 1, characterized in that that the regulator is N-ethyl malmide, iodoacetic acid, iodosobenzoic acid or iodoacetamide used. 6. The method according to claim 1, characterized in that there is used as a regulator DL-isoleucine or a yeast preparation with a content of-amino acids . 7. The method according to claim 1, characterized in that the regulator nicotinic acid or its amide, riboflavin, thiamine hydrochloride, D-calcium panthothenate, choline chloride, acetylcholine chloride, folic acid, meso-inositol, pyridoxal chloride, p-aminobenzoic acid, substances in Yeast, cotton seed, cereal or soybean extracts that are rich in B vitamins, or the acetate of vitamin A, the acetate of DL-α-tocopherol or rutin are used. S. The method according to claim 1, characterized in that the regulator used is glycine, DL-allylglycine, «-picolinic acid hydrochloride, oxythiamine hydrochloride, DL-desthiobiotin or deoxypyridoxine hydrochloride. 9. The method according to claim 1, characterized in that the regulator used is chloramphenicol, Griseofulvin or the sodium salt of penicillin G is used. 10. The method according to any one of claims 1 to 9, characterized in that the regulator in an amount up to about 1,000 parts by weight per 1,000,000 parts by weight of air-dry plant material used. 11. The method according to any one of claims 1 to 10, characterized in that the plant material is previously treated with a solvent for saponins extracted and the extract is then acid hydrolyzed with cleavage of the glycosidic bond would. 12. The method according to any one of claims 1 to 11, characterized in that the vegetable material previously under hydrolysi he ends for Cleavage of the glycoidic bond is treated and then treated with a solvent for sapogenins was extracted. 13. The method according to claim 11 or 12, characterized characterized in that the treatment with the regulator is carried out before the hydrolysis. 14. The method according to any one of claims 1 to 13, characterized in that the vegetable material was incubated in the presence of an aqueous medium. 15. The method according to claim 14, characterized in that the incubating mass so A lot of water was added so that an aqueous phase was present during the entire incubation period. 16. The method according to claim 14 or 15, characterized characterized in that the regulator is added at the beginning of or during the incubation period. 17. The method according to any one of claims 1 to 16, characterized in that the vegetable material prior to obtaining the saponin or sapogenin with a steroid precursor in Was brought into contact. 18. The method according to claim 17, characterized in that as a precursor of a steroid Mevalonic acid, a derivative of this acid, or squalene was used. 19. The method according to any one of claims 1 to 16, characterized in that the vegetable material was brought into contact with a saturated hydrocarbon with 10 to 36 carbon atoms in the molecule before the extraction of the saponin or the sapogenin. 20. The method according to claim 19, characterized in that n-hexadecane, n-pentacosane or n-octacosane was used as the saturated hydrocarbon. 21. The method according to claim 19, characterized in that squalane was used as the saturated hydrocarbon. 22. The method according to any one of claims 1 to 21, characterized in that a starting material is used from the tubers of Dioscorea, the fruits of Solarium or Balanites or the seeds of Balanites or Trigonella was won.